Trench Fever in German Mass Burial

Trench fever seems to be all the rage these days in paleomicrobiology. It seems as though every time Bartonella quintana is added to a panel of pathogens for aDNA screening its found at some level. So far its been found in in a tooth from 4000 before present, in late medieval Venice, 14th century France, and Napoleonic Europe.

Construction at the University of Kassel in Germany discovered a mass grave  revealing “most of the individuals had been males of the age classes juvenis and adultas“. Grumbkow et al (2011) report that local historians, anthropologists and medical examiners concluded that it was a military cohort that died of epidemic disease in the late 18th to early 19th century. They focused on a reputed outbreak of typhoid fever in the winter of 1813/14 that had been linked with regional epidemics started by Napoleon’s troops fleeing from the battle of Leipzig. However, Grumbkow et al (2011) do not present any evidence to directly link their sample of remains to any army that took part in the battle, nor the reasoning behind dating the remains to late 18th to early 19th century for that matter.

To investigate whether these people were victims of a typhoid fever outbreak, Grumbkow et al obtained samples of 18 skeletons for DNA screening. They note that typhoid fever was a common diagnosis for any fever that caused red spots, but could include typhus, parathyroid fever and trench fever. Therefore they screened DNA extracted from 16 femurs and 2 humeri for Salmonella species, Salmonella enterica typhi, Bartonella quintana, Rickettsia prowazekii, and for the bacterial 16S rDNA (ribosomal DNA). As you might have guessed, they only found Bartonella quintana in three bones, that is 16.6.%. This is roughly what Roualt et al (2006) found at Vilnius at 20% for Bartonella quintana.

However, I do have a few concerns with this paper. First, the work is incomplete. They mention in their discussion that they need to screen for more Bartonella quintana genes to confirm their results. This is especially important because the samples they did amplify were all 100% identical to the genebank sequences (making contamination more likely). They list the 16S rDNA primers along with the others but never present any data for these primers. They used diluted “bacterial-positive control DNA” as a control for PCR inhibitors from the soil, which may be what the 16S rDNA primers were used for but it is not explicit. Contamination concerns usually preclude the use of positive controls but they write that “amplification with positive controls and spiked samples always showed the expected results”. The near standard ‘suicide PCR’ protocol for aDNA was not used. Brief communication or not, more information is needed.

Grumbkow, P., Zipp, A., Seidenberg, V., Fehren-Schmitz, L., Kempf, V., Groß, U., & Hummel, S. (2011). Brief communication: Evidence of Bartonella quintana infections in skeletons of a historical mass grave in Kassel, Germany American Journal of Physical Anthropology DOI: 10.1002/ajpa.21551  [Epub ahead of print]